3), this difference remained significant [uANOVA, F(1,148) = 730.1; P < 0.001]. Considering that activity in the center has been largely used as an indicator of anxiety ( Prut and Belzung, 2003), the ambulation in the center was analyzed separately. Regarding total ambulation (C + Pe), treatment with vinpocetine significantly ameliorated the hyperactivity induced by early ethanol exposure in a dose-dependent way [rANOVA: Neonatal Dasatinib concentration Treatment × Treatment at P30 interaction, F(2,63) = 3.6; P < 0.05]. As depicted in Fig. 1, the ambulatory activity of the ETOH + DMSO group was ∼29% higher than that of the SAL + DMSO

group (FPLSD, P < 0.05), ∼45% higher than that of the SAL + Vp10 mg group (FPLSD, P < 0.05) Talazoparib cost and ∼49% higher than that of the ETOH + Vp20 mg group (FPLSD, P < 0.01). The dose-dependent amelioration of hyperactivity elicited by vinpocetine was evidenced by the fact that the ETOH + Vp20 mg group had an average locomotor activity similar to that of the SAL + DMSO group while, distinctively, the ETOH + Vp10 mg group did not differ from both the SAL + DMSO and the ETOH + DMSO groups. No significant differences were observed between SAL + Vp20 mg and ETOH + DMSO as well as between males and females (P > 0.05 in all pairwise comparisons). For both ambulation in the center and C/Pe ratio data, increases in values were observed along the 10 time-intervals [rANOVA: ambulation in the center, F(6.3,393.1) = 3.3; P < 0.01 and

C/Pe ratio, F(3.6,120.1) = 2.7; P < 0.05]. However, for these two variables, no differences were observed between groups. Furthermore, no effects or interactions regarding gender, neonatal exposure and treatment at P30 were observed. Taken together, these results suggest that the ethanol-injected mice are hyperactive while maintaining normal levels of anxiety. In addition, the treatment with vinpocetine did not differentially affect the anxiety

levels of ethanol- or saline-injected animals. Regarding ambulation in the periphery, the results were similar to those described for total ambulation (C + Pe) (Supplementary Material, B). Considering that the vinpocetine treatment effectively ameliorated Tryptophan synthase hyperactivity only at the 20 mg/kg dose, we did not conduct the cAMP assays on the vinpocetine 10 mg/kg samples. As expected, treatment with vinpocetine increased the levels of cAMP by approximately 60% both in the hippocampus [uANOVA: F(1,21) = 69.8; P < 0.001] and in the cortex [uANOVA: F(1,21) = 43.8; P < 0.001]. In the hippocampus, neonatal exposure to ethanol reduced cAMP levels [uANOVA, F(1,21) = 63.9; P < 0.001] and treatment with vinpocetine significantly restored cAMP levels [uANOVA, F(1,21) = 9.1; P < 0.01]. Accordingly, cAMP levels in the ETOH + DMSO group were significantly lower than those observed in both SAL + DMSO (∼33%) and ETOH + Vp20 mg (∼31%) groups, which, in turn, did not differ from each other ( Fig. 2A). No significant differences were observed between males and females.